Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
  • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
  • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
  • C07K16/2803—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
  • C07K16/2833—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily against MHC-molecules, e.g. HLA-molecules
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
  • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
  • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
  • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
  • A61K47/6835—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site
  • A61K47/6851—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site the antibody targeting a determinant of a tumour cell

Definitions

• the invention relates to methods of producing medicaments for combatting tumors and to the use thereof for said purpose.
• the chemical or biological tumor therapy has not yet focused on individual tumor cell types of a single patient but still detects in general all rapidly dividing cells, independent of whether or not they are malignant and independent of the individual patient.
• the basis for the well-regulated existence of an entire organism is the communication between the cells or the cellular dialogue.
• This dialogue and its regulation enable to maintain the existence of an entire organism even though cells constantly die and/or are reproduced.
• the differentiation of cells is also regulated, as known from stem cell research.
• This dialogue even enables the well-regulated cooperation of two different cell clones even if one shows an extremely fast growth as is the case during pregnancy.
• the basis of the cellular dialogue in man is the MHC (major histocompatibility complex) with its HLA groups.
• the identification of a cell by HLA groups is the basis of every cellular communication.
• the cellular communication in which develops in cooperation with specific receptors, such as the killer-immunoglobulin-like receptors (KIR) on the natural killer cells (NK cells) or the LILR (leukocyte immunoglobulin-like receptors), wit subsequent involvement of further factors, such as cytokines, growth factors, etc.
• KIR killer-immunoglobulin-like receptors
• NK cells natural killer cells
• LILR leukocyte immunoglobulin-like receptors
• HLA groups D (DR, DP, DQ, etc.; MHC II): They play an important role in immunocompetent cells and/or in the antigen presentation.
• HLA groups E, F and G They identify embryonic cells, in particular on the so-called front of invasion.
• HLA group H and the following: The function of these so-called “pseudo-antigens" is not yet quite clear.
• the MHC complex also comprises further substances such as the complement factors which belong to class III.
• a tumor cell basically has a genetic code the same as that of any other cell of the entire organism. Therefore, it does not have any information other than that of the entire organism with respect to cell division and cell differentiation. As a result, every malignant tumor disease is unique and individual, i.e. specific to the respective organism.
• oncogenes can also form an integral part of the genetic material from procreation on. Oncogenes and/or the activation thereof and other external factors can permanently affect the biology of a tumor cell. Yet, the tumor cells stay involved in the cellular dialogue of the entire organism and the regularities valid therein.
• Organisms having a high cellular differentiation such as human beings, "pay” for their high differentiation by a loss of multipotency or totipotency.
• a loss of multipotency or totipotency In the case of organ losses, restitutio ad integrum is no longer possible but only the repair by connective tissue.
• the loss of totipotency or multipotency is not so distinct and therefore when an arm is lost, for example, a new arm can grow again even though it is smaller.
• Totipotency is basically encoded in the genetic material of higher living beings as well. This is proved by the simple fact that this genetic material formerly had to control the development from a fertilized egg cell to a differentiated organism. Cloning experiments also show that "resetting" of the genetic material ("reprogramming") of even highly differentiated cells, such as the udder cells of the cloned sheep "Dolly", "to zero” is possible in the nucleus. In the final analysis, this also applies to the procreation and/or fertilization of an egg cell where the genetic material of two relatively old individuals (father and mother to be) is reset "to zero" and is encoded again for the development of a new living being.
• a tumor cell is also provided with genetic material that fundamentally codes for all growth and differentiation processes which are at all possible in an entire organism, i.e. also for the mechanisms of the initial embryonic implantation, of the early embryo-maternal cell dialog and of the subsequent embryonic-fetal development.
• Essential characteristics of this "way back” are the loss of cellular differentiation and the loss of specific cell performances and also the (re)gaining of uncontrolled cell growth.
• tumor cells can express typical embryonic HLA groups on their surface.
• embryonic HLA groups namely the known ones and optionally also not yet identified ones, e.g. in the scope of so-called "pseudogenes”
• the expression of these typical HLA groups on the surface enables cells to activate corresponding receptors e.g. on the NK cells but also the lymphocytes and further immunocompetent cells, and therefore there is not only no attack of the unspecific immune defense, i.e. the NK cells and lymphocytes, but also in the individual case tumor cells (and also embryonic cells) are able to "let the immune defense work for them", namely by a synthesis of growth factors and cytokines which are beneficial to the own development.
• TAM tumor-associated macrophages
• MDSC tolerogenic "myeloid-derived suppressor cells”
• cytokines such as MIF (macrophage migration inhibition factor) which is likely to be produced in the tumor (likely by NK cells) and has a proangiogenetic effect, thus supporting the proliferation and migration of tumor cells.
• Tumor cells need not be very resistant. As is known, they are more sensitive to chemotherapy and also more sensitive to radiation than "healthy" and differentiated standard cells. Their cell division rate is not particularly high either. The danger resulting from a malignant tumor cell is above all that it is able on the basis of the cellular communication to "enforce" the progressing uncontrolled growth.
• malignant stem cells spread and colonize. Should this be correct, such "malignant stem cells” should by means of the cellular communication with the adjacent tissue also be locally able to evade the growth control and the differentiation pressure. Even if formed from a dedifferentiation, stem cells would behave like stem cells in general, which in this case directs the focus in particular on the mode of functioning of the embryomaternal communication (of the embryomaternal dialogue).
• the malignant degeneration of a cell is a unique process which is specific to every individual. This is not altered by the fact that there are pathologically well classifiable (always recurring) tumor types across individuals. This circumstance is rather a proof for the fact that a malignant tumor does not form by every dedifferentiation and every "way back". It is rather likely that only certain constellations can "survive” on the "way back", thus resulting in the typical tumor entities across individuals.
• the dedifferentiation or "degeneration" of a cell is presumably a comparatively ubiquitous process in every entire organism. However, it almost never leads to the formation of a tumor disease since only some few of these cells have the cell-biological and also communicative preconditions (from cell to cell) that are necessary for the survivability.
• the cells which show survivability use the two above mentioned mechanisms, presumably in a combined form.
• the "acquired” immunity develops during pregnancy and means that there is not only a tolerance with respect to the body's own HLA groups but also always with respect to the foreign adult HLA groups of one's own mother.
• a tumor cell basically expresses the same adult HLA groups as all other somatic cells of the entire organism. As a result, it is protected from an attack of the specific immune defense. This also applies in principle when parts of the original HLA pattern are lost "on their way back", are expressed less densely or are available in a changed, i.e. corrupted, form (which is not atypical for tumor cells). It is here useful to make a comparison with an embryo's embryonic cells which were flushed out in the maternal organism and persist therein (this is referred to as "microchimerism).
• the embryonic surface structures, in particular HLA-G, -E and -F, on the embryonic cells prevent the mother's immune system from attacking the cells.
• the embryonic cells are presumably not attacked throughout the mother's life - similar to tumor cells.
• This inflammatory counter-reaction of the organism does not take place in the case of tumor cells since the tumor cells form HLA characteristics on the surface during further differentiation, said characteristics not differing from those of the host organism.
• This complete or incomplete expression of the adult HLA patterns prevents an attack of the specific immune defense even when the tumor cell stands out by additional antigen expressions (or overexpressions) as known and described for tumor cells.
• the protection resulting from the complete or incomplete expression of the original adult HLA patterns is obviously very effective such that a tumor cell can express its specific antigen patterns (thus "standing out” as a result) without an effective attack of the (specific) immune response, i.e. also of the B lymphocytes and T lymphocytes occurring. It is remarkable that the tumor antigen expression patterns are relatively specific to individual tumor types.
• MHC-/HLA groups make the antigen presentation cascade (APM (antigen processing machinery)) more and more faulty and therefore typical, human-associated or own antigens are hardly presented or are not presented (any more).
• APM antigen processing machinery
• monocytes and the macrophages resulting from them also play a role in tumor growth.
• macrophages can only be activated when cells of the unspecific immune defense (such as NK killer cells) or the specific immune defense (such as T-cells or B-cells) are present.
• NK killer cells such as NK killer cells
• specific immune defense such as T-cells or B-cells
• this requires "priming” where antigen-presenting cells (such as the dendritic cells) present mutated or "foreign” proteins, thus leading to the formation of cytotoxic T-cells.
• numerous cytokines such as interferon ⁇ (IFN- ⁇ ) or the tumor necrosis factor (TNF- ⁇ ) play a role as well.
• IFN- ⁇ interferon ⁇
• TNF- ⁇ tumor necrosis factor
• Macrophages can be found in the basal endometrium in the case of an establishing pregnancy. They usually have an inhibitory effect on the invasion behavior of the embryo and form so to speak a "protective wall" between the implanting embryo and the myometrium.
• the embryo secretes macrophage migration inhibiting factors, i.e. factors which limit and inhibit the attack of the macrophages. This applies likewise to malignant tumors (see above)
• the invention in a first aspect, relates to a method of producing a medicament for combatting a tumor in a host, comprising determining the individual embryonic HLA expression pattern of a tumor cell of a specific tumor, including in particular HLA-E, HLA-F, HLA-G and others and optionally pseudoantigens, developing antibodies against the expression pattern and coupling the antibodies with a cytotoxin, such as a chemotherapeutic agent or a radioisotope, wherein the thus created individual antibodies serve as an antitumor medicament and/or the antibodies are used to destroy or mask the tumor-specific embryonic HLA pattern.
• a cytotoxin such as a chemotherapeutic agent or a radioisotope
• the invention relates to the use of a medicament which is produced as described in the first aspect for combatting a tumor.
• the invention in a third aspect, relates to a method of producing a medicament for combatting a tumor in a host, comprising removing tumor cells with an individual expression pattern and exposing them in vitro to immunocompetent cells, preferably of the host, thus "training" the immunocompetent cells against the individual expression pattern and using the thus trained cells as an antitumor medicament for the in vivo introduction into the host.
• the invention relates to the use of a medicament which is produced as described in the third aspect for combatting a tumor.
• the invention in a fifth aspect, relates to a method of producing a medicament for combatting a tumor in a host, wherein heterologous tumor cells are selected which have tumor-specific antigen patterns the same or similar to those of the tumor cells of the host, but have adult HLA groups completely differing therefrom (and - if possible - from the mother), the selected heterologous tumor cells are freed, i.e. are denuded, from the embryonic HLA expression pattern by means of a medicament produced according to the first aspect, and killed and the resulting heterologous tumor cells serve as an antitumor medicament for introduction into a host.
• the invention relates to the use of a medicament which is produced as described in the fifth aspect for combatting a tumor.
• the invention relates to a method of producing an antitumor antibody, wherein with respect to a host heterologous tumor cells are selected which have tumor-specific antigen patterns the same or similar to those of the tumor cells of the host, but have adult HLA groups fully differing therefrom (and from the mother), the selected heterologous tumor cells are freed, i.e.
• the invention relates to the use of the antibody produced by the method of the seventh aspect as an antitumor medicament.
• the invention relates to a method of producing genetically modified immunocompetent cells, in particular genetically modified NK cells and also genetically modified lymphocytes, comprising silencing the genes for the receptors which convey an inhibitory effect and are activated by the tumor's own HLA groups, whether they are adult (MLA-A, -B, -C, -D) or embryonic (HLA-E, -F, -G inter alia ).
• the cells can be proliferated in vitro.
• the invention relates to immunocompetent cells which lack the receptors conveying an inhibitory effect which are activated by the tumor's own HLA groups, whether they are adult (MLA-A, -B, -C, -D) or embryonic (HLA-E-, - F, -G inter alia ).
• NK cells are concerned here whose genes for the receptors which convey an inhibitory effect and which recognize the body's own HLA-A, -B, -C and/or -D are deleted.
• the invention relates to the use of the cells according to the tenth aspect as a medicament for combatting a tumor.
• a first aspect of the invention relates to a method of producing a medicament for combatting a tumor in a host, comprising determining the individual embryonic HLA expression pattern of a tumor cell (of a specific tumor), in particular HLA-E, HLA-F, HLA-G and others and optionally pseudoantigens, developing antibodies against the expression pattern and coupling the antibodies with a cytotoxin, such as a chemotherapeutic agent or a radioisotope (favorably an ⁇ -radiation source).
• a cytotoxin such as a chemotherapeutic agent or a radioisotope (favorably an ⁇ -radiation source).
• a second aspect of the invention relates to the use of a medicament which is produced as described in the first aspect for combatting a tumor.
• the antibodies destroy or mask tumor-specific embryonic HLA patterns.
• the partial or full recovery of embryonic HLA expression patterns on malignant cells can serve for identifying and combatting these malignant cells.
• HLA groups F and G are predominantly the HLA groups F and G but also groups E and others and possibly also some pseudoantigens, i.e. HLA groups H and higher.
• HLA groups H and higher it is characteristic of these embryonic HLA groups that they are not only able to interact e.g. with specific receptors on NK cells (NKG2-/KL (killer lecithin-like receptors) or the KIR formations or the lymphocytes (LIL receptors) and/or immunoglobulin-like transcripts (ILT) but that these interactions besides a "blockade” of the immune system also lead as to the fact that various immunocompetent cells (e.g.
• NK cells and macrophages synthesize and secrete cell growth- and cell division-promoting cytokines and growth factors.
• the tumor cell to manipulate the cellular dialogue so as to enable unlimited cell growth.
• the secreted cytokines and growth factors support this uncontrolled cell growth and/or cell division behavior.
• stem cells which usually do not express any adult HLA groups (A to D) and therefore are especially a target of the unspecific immune defense, i.e. the NK cells and the macrophages, this mechanism is of major significance.
• chemotherapeutic agents or radioisotopes it is possible to couple e.g. chemotherapeutic agents or radioisotopes to such antibodies to thus damage the tumor cell.
• a second possibility is to destroy the embryonic HLA patterns in this way, e.g. by combining the antibodies with enzyme complexes which directly attack HLA antigens or at least the scaffold proteins of the HLA complex. Furthermore, it would also be possible to "mask" the entire embryonic HLA complex by means of these antibodies such that it can no longer be recognized as such by the immune defense. As a result, the above described mechanisms of the "dialogue” would be omitted.
• the implementation of the described measures does not have to take place in vivo. It is also basically possible to remove tumor cells and to "train" immunocompetent cells - after removal or masking of the embryonic HLA structures -by means of them in vitro.
• a third aspect of the invention relates to a method of producing a medicament for combatting a tumor in a host, comprising removing tumor cells with an individual expression pattern and exposing them in vitro to immunocompetent cells, preferably of the host.
• the immunocompetent cells are trained against the individual expression pattern, and the thus trained cells serve as an antitumor medicament for in vivo introduction into the host.
• a fourth aspect of the invention is the use of a medicament which is produced as described in the third aspect for combatting a tumor.
• the “in vitro training” offers the advantage of "training” the individual cell types of the unspecific and specific immune defense in controlled fashion by means of the specific HLA and tumor antigen expression patterns so as to be able to better dose the effect when these cells are returned to the original organism.
• the typical embryonic expression patterns must be removed or “masked” in vitro , but this also applies to the adult expression patterns (HL groups A to C and/or D and the possibly existing HLA groups which correspond to those of the own mother), since the immunocompetent cells of the specific immune defense, such as T-lymphocytes, can hardly or only poorly attack cells which have the typical adult expression patterns of the original organism.
• the in vitro "training" of the immunocompetent cells of the specific and unspecific immune defense serves predominantly for training them against the individual-specific tumor antigen expression patterns -and this after the discontinuation of the blockade by the embryonic and adult HLA groups.
• the adult HLA patterns of the original organism which are also fully or partially expressed by the tumor cells, thus represent the second effective protection of a tumor cell against an attack of the body's own immune defense, namely the cells of the specific immune defense.
• the invention thus relates to a method of producing a medicament for combatting a tumor in a host, wherein heterologous tumor cells are selected which have tumor-specific antigen patterns that are equal or similar to those of the tumor cells of the host but have adult HLA groups completely differing therefrom, the selected heterologous tumor cells are freed, i.e. denuded, from the embryonic HLA expression pattern by means of a medicament produced according to the first aspect, and are killed and the resulting killed heterologous tumor cells serve as an antitumor medicament, optionally together with an adjuvant, e.g. aluminum hydroxide or an oil-in-water emulsion, which comprises squalene, polysorbate 80 and vitamin E, for introduction into the host.
• an adjuvant e.g. aluminum hydroxide or an oil-in-water emulsion, which comprises squalene, polysorbate 80 and vitamin E
• a sixth aspect of the invention is the use of a medicament which is produced as described in the fifth aspect and optionally contains an adjuvant, for combatting a tumor.
• Tumor cells which in the model case have a tumor-specific antigen expression pattern the same as that of the host organism however fully differ therefrom as regards the HLA groups, induce an immune response against these tumor cells in the host, namely on account of the divergent HLA expression patterns.
• the specific immune defense will also "learn" the tumor antigen expression patterns and form antibodies and cells of the cellular immune defense which are directed thereagainst.
• the immunological response i.e. the formation of antibodies and/or the cellular immune response to the tumor-specific antigen expressions, can be promoted by adjuvants, such as aluminum hydroxide or an oil-in-water emulsion which comprises squalene, polysorbate 80 and vitamin E.
• the immune response can also be evoked in vitro ( in vitro "training" of the immunocompetent cells).
• the antibodies and the trained immunocompetent cells can then be used in vivo or in vitro to attack the individual-specific tumor cells.
• the adult HLA groups (A to D) which are fully or partially left on the surface protect a tumor cell against an attack of the specific immune defense.
• the immune defense In order to carry out a sensitization of the immune defense against the typical surface expression patterns of a tumor cell, it is thus necessary to supply killed tumor cells with oncogenes as identical as possible and an HLA expression pattern as divergent as possible.
• the difficulty is to identify tumor cells being as HLA-A to D-divergent as possible and having tumor-specific antigen patterns as equal as possible (as in the host organism). While it is relatively easy to determine the HLA patterns, this is markedly more difficult with the typically tumor-specific antigen expressions. Here, only a "constriction" can be made for the time being, namely by means of the greatest possible number of different (known) antigen expression patterns. Thus, there is a residual insecurity which can, however, be avoided by basing the therapy on different tumor cell lines having HLA patterns as discrepant as possible and tumor antigen expression patterns as similar as possible.
• heterologous tumor cells it is also necessary for the heterologous tumor cells not to have any embryonic HLA groups and/or features; therefore, they also have to be denuded.
• the invention further relates to a method of producing an antitumor-antibody, wherein with respect to a host heterologous tumor cells are selected which have tumor-specific antigen patterns equal or similar to the tumor cells of the host, but have adult HLA groups which fully differ therefrom, the selected heterologous tumor cells are freed, i.e.
• An eighth aspect relates to the use of the antibody produced by the above method as an antitumor medicament.
• a ninth aspect of the invention relates to a method of producing genetically modified immunocompetent cells, in particular NK cells, comprising silencing the genes for the receptors which convey an inhibitory effect and are activated by the tumor's own HLA groups.
• NK cells genetically modified immunocompetent cells
• a tenth aspect of the invention relates to the thus produced immunocompetent cells, in particular NK cells.
• An eleventh aspect of the invention relates to the use of the cells of the tenth aspect as a medicament for combatting a tumor.

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• 2011
  • 2011-08-25 DE DE102011111631A patent/DE102011111631A1/de not_active Withdrawn
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